Visualizing the Dynamics of Inactive X Chromosomes in Living Cells Using Antibody-Based Fluorescent Probes

  • Yuko Sato
  • Timothy J. Stasevich
  • Hiroshi KimuraEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1861)


The inactive X chromosome (Xi) harbors characteristic epigenetic features, including the enrichment of histone H3 lysine 27 trimethylation (H3K27me3) and H4 lysine 20 monomethylation (H4K20me1) as well as a lack of histone acetylation. Recently, these modifications have been visualized not only in fixed specimen, but also in living cells via probes derived from modification-specific antibodies. The probes include fluorescently labeled antigen binding fragments (Fabs), which can be loaded into cells, as well as genetically encoded single-chain variable fragments tagged with the green fluorescent protein. We refer to the latter as modification specific intracellular antibodies, or “mintbodies” for short. By using Fabs or mintbodies to target Xi-specific modifications, the dynamics of Xi in living cells can be visualized.

Key words

Histone modifications Live-cell imaging Mintbody Monoclonal antibody 



This work was supported by JSPS KAKENHI Grants JP15K07157 (to Y.S.) and JP25116005, JP26291071, and JP17H01417 (to H.K.).


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Yuko Sato
    • 1
  • Timothy J. Stasevich
    • 2
    • 3
  • Hiroshi Kimura
    • 1
    • 2
    Email author
  1. 1.Cell Biology Center, Institute of Innovative ResearchTokyo Institute of TechnologyYokohamaJapan
  2. 2.World Research Hub Initiative, Institute of Innovative ResearchTokyo Institute of TechnologyYokohamaJapan
  3. 3.Department of Biochemistry and Molecular BiologyColorado State UniversityFort CollinsUSA

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